Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H5/00—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium
  • C07H5/04—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium to nitrogen
  • C07H5/06—Aminosugars
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
  • C07H15/02—Acyclic radicals, not substituted by cyclic structures
  • C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
  • C07H15/10—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical containing unsaturated carbon-to-carbon bonds
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
  • C07H15/18—Acyclic radicals, substituted by carbocyclic rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
  • C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H7/00—Compounds containing non-saccharide radicals linked to saccharide radicals by a carbon-to-carbon bond
  • C07H7/02—Acyclic radicals
  • C07H7/027—Keto-aldonic acids

Definitions

• This invention relates to sialic acid derivatives and galactose derivatives, more particularly gangliosides and intermediate compounds for use in producing gangliosides, and to methods for producing the same.
• Glycolipids found in mammal cells are glycosides between ceramides, which are sphingosines (long chain amino alcohols) to which aliphatic acids have been attached through an amide linkage, and one or more sugars such as glucose, galactose, N-acetyl glucosamine, N-acetyl galactosamine, fucose, sialic acid, etc.
• ceramides which are sphingosines (long chain amino alcohols) to which aliphatic acids have been attached through an amide linkage
• sugars such as glucose, galactose, N-acetyl glucosamine, N-acetyl galactosamine, fucose, sialic acid, etc.
• gangliosides those containing sialic acid are called gangliosides.
• Gangliosides exist mainly in the outer molecular layer of double molecular layers of the mammal cell membrane. Recent studies show that gangliosides play important roles in reception and recognition of, and response to, information in cells, receptor mechanism, differentiation, cell propagation, malignant cell transformation, cell behavior, etc.
• sialic acid residue-containing oligosaccharides it is very difficult to isolate sialic acid residue-containing oligosaccharides from an organism. Therefore precise synthesis of such sialic acid residue-containing oligosaccharides is necessary for the elucidation of the precise correlation between biological information and the molecular structure of the oligosaccharides.
• An object of this invention is to provide novel gangliosides, novel sialic acid derivatives for use in producing the gangliosides and methods for producing the same.
• Another object of this invention is to provide novel galactose and lactose derivatives which can be used as intermediate compounds in producing the gangliosides and methods for producing the same.
• novel sialic acid derivatives of this invention are represented by the formula I.
• the galactose derivatives of this invention are represented by the formula:
• galactose derivatives provided by this invention are also lactose derivatives.
• Ceramide moieties of gangliosides can be prepared by the method as shown in Scheme lb.
• Compound (I) can be prepared by the method as shown in Scheme la (see the specification of Japanese Patent Application No. 59-44913).
• Compound (ii) is obtained by refluxing overnight an alkyl halide such as 1-bromotetradecane and triphenylphosphine in a solvent such as xylene.
• 1,2-O-isopropylidene- ⁇ -D-xylo-pentodialdo-1,4-furanose (i) is reacted with the compound (ii) in a solvent, e.g. tetrahydrofuran (THF) and hexane in the presence of BuLi to obtain 4-alkylvinyl derivative (iii).
• a solvent e.g. tetrahydrofuran (THF) and hexane in the presence of BuLi to obtain 4-alkylvinyl derivative (iii).
• the reaction temperature and time are preferably in the range of -15°C to 25 0 C and 0.5 to 24 hours, respectively.
• the compound (iii) is treated with methanesulfonylchloride in dry pyridine to obtain 3- methanesulfonyl derivative (iv).
• the reaction temperature and time are preferably in the range of 0°C to 25°C and 2 to 24 hours, respectively.
• the compound (iv) is treated in acetic acid - water to remove isopropylidene group.
• Diol derivative (v) is obtained.
• the reaction temperature and time are preferably in the range of 70 0 C to 90°C and 0.5 to 5 hours, respectively.
• the compound (v) is treated with an oxidizing agent (e.g. sodium metaperiodate) in a solvent (e.g. ethanol) to cleave the diol and then treated with a reducing agent (e.g. sodium borohydride) to obtain diol (vi).
• an oxidizing agent e.g. sodium metaperiodate
• a solvent e.g. ethanol
• a reducing agent e.g. sodium borohydride
• the oxidation reaction is preferably conducted at 0°C to 25°C for 0.5 to 24 hours.
• the reduction reaction is preferably conducted at 0°C to 10°C for 0.5 to 2 hours.
• the compound (vi) is reacted with an alkyl vinyl ether such as ethyl vinyl ether in a solvent such as dichloromethane in the presence of a catalyst such as p-toluenesulfonate to obtain di-alkyl vinyl ether (vii).
• This reaction is preferably conducted at 0°C to 30 0 C for 0.5 to 24 hours.
• the compound (vii) is treated with an azide such as sodium azide in a solvent such as dimethylformamide (DMF) to obtain azide (viii).
• This reaction is preferably carried out at 70oC to 1200C for 8 hours to six days.
• the azide (viii) is reduced by a reducing agent such as sodium borohydride and Lindler catalyst/H 2 in a solvent such as ethanol and isopropanol to give amine (ix).
• a reducing agent such as sodium borohydride and Lindler catalyst/H 2 in a solvent such as ethanol and isopropanol
• the reaction is carried out at a reflux temperature for one to six days when sodium borohydride is used and at 0°C to 30°C for 2 to 24 hours at a hydrogen pressure of 1 to 4 atms. when Lindler catalyst/H 2 is used.
• the amine (ix) is reacted with an acyl halide in the presence of an amine such as pyridine and dimethylaminopyridine to obtain amide (x) or (xi). This reaction is preferably carried out at 0°C to 30°C for 0.5 to 24 hours.
• the amine (ix) dissolved in a solvent such as dichloromethane is reacted with an aliphatic acid in the presence of 2-chloro-l-methylpyridinium iodide and tri-n-butylamine to obtain the amide (x) or (xi).
• This reaction is preferably carried out at a reflux temperature for 0.5 to 13 hours under an inert atmosphere.
• the amide (x) or (xi) is treated with pyridinium p-toluenesulfonate, Amberlist A-15 (tradename), etc. in a solvent such as methanol or dichloromethane to obtain ceramide (xii) or (I).
• the compound (I) thus obtained is treated with trityl chloride in pyridine to obtain trityl derivative (II) which is then treated with benzoyl chloride and dimethylaminopyridine to obtain trityl - benzoyl derivative (III) which is then treated with p-toluenesulfonic acid to remove the trityl group.
• Benzoyl ceramide (IV) is obtained.
• the compound (IV) can be obtained without the isolation of the compounds (II) and (III).
• Lactose and galactose derivatives which can be used in producing gangliosides of this invention can be produced by the processes as shown in Schemes 2a and 2b.
• D-Lactose octaacetate (1) is treated with tri (n-butyl) tin allyloxide in a solvent such as ethylene chloride in the presence of a catalyst such as tin tetrachloride to obtain allyl derivative (2) which is then deacetylated by a conventional manner, e.g. by NaOMe/MeOH to obtain the deacetylated compound (3) which is further reacted with 2,2-dimethoxypropane and p-toluenesulfonic acid in acetone/DMF.
• 3 1 , 4'-O-isopropylidene derivative (4) and 4', 6 1- 0-isopropylidene derivative (5) are obtained.
• the compound (4) is acetylated by acetic anhydride/pyridine, followed by the treatment with 90% CF 3 COOH to obtain the compound (11).
• Benzyl 3', 4'-O-isopropylidene lactose (F) is treated in DMF with benzyl bromide in the presence of NaH to obtain benzyl derivative (G) which is then treated with aqueous acetic acid solution to remove isopropylidene group. Hexa-O-benzyl derivative (H) is obtained.
• Galactose derivatives which can be used in producing the sialic acid derivatives of this invention can be prepared as follows. Benzyl galactoside (A) .suspended in acetone is reacted with 2,2-dimethoxypropane in the presence of p-toluenesulfonic acid to obtain 3,4-0-isopropylidene derivative (B) which is then reacted with benzyl bromide in a solvent, e.g. DMF in the presence of NaH to convert it into tribenzyl derivative (C) which is subsequently treated with aqueous acetic acid solution to remove isopropylidene group. The compound (D) is obtained.
• the reaction between the compound (8), (9), (11), (D) or (H) and the compound (E) is carried out in a solvent such as dichloromethane or 1,2-dichloroethane in the presence of a glycosidation catalyst such as Hg(CN) 2 , HgBr 2 , molecular sieve (hereinafter referred to as MS), Ag 2 C0 3 , AgC10 4 , AgOS0 2 CF 3 , (CH 3 ) 3 COS0 2 CF 3 , etc. at -20°C to 150°C for 1 to 120 hours.
• a glycosidation catalyst such as Hg(CN) 2 , HgBr 2 , molecular sieve (hereinafter referred to as MS), Ag 2 C0 3 , AgC10 4 , AgOS0 2 CF 3 , (CH 3 ) 3 COS0 2 CF 3 , etc. at -20°C to 150°C for 1 to 120 hours.
• Trisaccharide (12), (13), (14), (45), (46) or (47) or disaccharide (31), (32) or (33) is obtained.
• the removal of protective groups of these compounds gives the desired compounds, respectively.
• the compound (13) is acetylated by acetic anhydride-pyridine to give the compound (15) which is then treated with PdCl 2 and AcONa/AcOH to remove allyl group.
• the resulting compound (16) is treated with monochloroacetic anhydride-pyridine to obtain the compound (17).
• Catalytic reduction of the compound (17) and subsequent acetylation give the compound (18) which is then treated sodium acetate and thiourea in a solvent such as ethanol to give the compound (19).
• ganglioside GM 3 (the compound (22)) may be produced as follows:
• These new compounds of this invention may be employed as tumor markers, differentiation markers of cells' having differentiation potency, or useful intermediates for the synthesis of various gangliosides.
• the compound (1) (975 mg, 1.5 mmol) was dissolved in pyridine (15 ml). TrCl (625 mg, 2.25 mmol) was added. The mixture was stirred at 55°C for 4 hours and at room temperature for 24 hours. Benzoylchloride (315 mg, 2.25 mmol) and dimethylaminopyridine (183 mg, 1.5 mmol) were added and stirred at room temperature for 5 hours. The reaction mixture was concentrated in vacuo and the residue was dissolved in ethyl acetate. The solution was washed with water, dried on MgS0 4 and concentrated in vacuo. The residue was dissolved in CH 2 Cl 2 (10 ml).
• n-Bu 3 Sn-O-CH 2 CHCH 2 80.7g, 0.23 mol was dissolved in ethylene chloride (500 ml). Tin tetrachloride (31.0 ml) was added under ice-cooled condition. To this solution, there was added 250 ml of a solution of D-lactose octaacetate (1) (142g, 0.21 mol) in ethylene chloride. The reaction mixture was stirred at room temperature for 2.5 hours and then poured into a saturated KF solution. Insoluble products precipitated were filtered. The filtrate was washed with a saturated NaCl solution, dried on MgS0 4 and concentrated in vacuo. The residue was subjected to column chromatography (silica gel 2 kg) and eluted with toluene-ethylacetate (1:1) to give the compound (2) (85.8 g, 57.8%).
• the compound (2) (85.8 g, 0.127 mol) was dissolved in methanol (600 ml). N-NaOCH 3 solution (10 ml) was added and stirred at room temperature for 2 hours. Precipitated crystals were collected by filtration. The compound (3) (41.8 g, 86.2%) was obtained.
• the compound (6) (2.47 g, 2.8 mmol) was dissolved in 60 ml of 90% acetic acid solution in water and stirred at 60°C for 3 hours. The reaction mixture was concentrated in vacuo. The residue was recrystallized from ether - hexane to give the compound (8) (1.24 g, 52.6%) as needle crystals.
• the compound (7) (1.66g, 1.9 mmol) was dissolved in 30 ml of 90% acetic acid solution in water and stirred at 60 °C for 3 hours. The reaction mixture was concentrated in vacuo. The residue was recrystallized from chloroform-hexane to give the compound (9) (l.Olg, 67.6%).
• the compound (4) (2.llg, 5.0 mmol) was dissolved in acetic anhydride (15 ml) and pyridine (15 ml). The reaction mixture was stirred at room temperature for 24 hours and concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 250g) and eluted with 3.5% MeOH-containing chloroform to give the compound (10) (2.31g, 73.1%).
• the compound (10) (15.5g, 23.9 mmol) was dissolved in 90 % CF 3 COOH solution and stirred at room temperature for 20 minutes. The reaction mixture was concentrated in vacuo. The residue was dissolved in ethyl acetate. The solution was washed with NaHC0 3 solution and then saturated NaCl solution, dried on MgS0 4 and concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 300g) and eluted with 4% MeOH-containing chloroform to give the compound (11) (11.0g).
• the compound (13) (605 mg, 0.57 mmol) was dissolved in pyridine (10 ml) and acetic anhydride (10 ml). To this solution, there was added dimethylamino-pyridine (70 mg). The mixture was stirred at room temperature for 24 hours and then concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 80 g) and eluted with toluene-ethylacetate (1:2) to give the compound (15) (452 mg, 72.6%).
• the compound (15) (410 mg, 0.34 mmol) was dissolved in 90% AcOH. To this solution, there were added AcONa (500 mg) and palladium chloride (540 mg). The mixture was stirred for 2 hours in an ultrasonic stirrer and then concentrated in vacuo. The residue was dissolved in ethylacetate. The solution was washed with water, dried on MgS0 4 and concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 50g) and eluted with 10% MeOH-containing isopropyl ether to give the compound (16) (353 mg, 89%).
• the compound (16) (312 mg, 0.24 mmol) was dissolved in pyridine (7 ml). Dry monochloroacetic acid (312 mg) was added and stirred at room temperature for one hour, to which ethylacetate was added to dilute it. The solution was washed with saturated NaHC0 3 solution, diluted HC1 and saturated NaCl solution, dried on MgS0 4 and concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 40g) and eluted with 10% MeOH-containing isopropyl ether to give the compound (17) (257 mg, 77.6%).
• the compound (17) (91 mg, 0.065 mmol) was dissolved in methanol (3 ml). 10 % Pd/C (50 mg) was added and catalytic reduction was carried out at room temperature for 24 hours. The reaction mixture was filtered to remove Pd/C and the filtrate was concentrated in vacuo. To the residue, there were added acetic anhydride (1.0 ml) pyridine (1.0 ml). The mixture was stirred at room temperature for 2.5 hours and concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 10 g) and eluted with 4% MeOH-containing chloroform to give the compound (18) (61 mg, 81%).
• the compound (18) (61 mg, 0.053 mmol) was dissolved in ethanol. To this solution, there were added thiourea (20 mg) and sodium acetate (4 mg). The mixture was heated and refluxed for 5 hours. The reaction mixture was concentrated in vacuo. The residue was dissolved in 4% MeOH-containing chloroform and subjected to silicagel column chromatography (Wakogel C-300, 10 g) to give the compound (19) (21 mg). The compound (18) (40 mg) was recovered.
• the compound (19) (20 mg, 0.0187 mmol) was dissolved in methylene chloride (0.5 ml) and trichloroacetonitrile (13.5 mg) was added. To this, there was added NaH (60% in oil) (1.0 mg) under ice-cooled condition and stirred for 2 hours. The mixture was concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 5 g) and eluted with ethyl acetate to give the compound (20) (10.0 mg).
• the compound (21) (4.0 mg) was dissolved in a mixed solvent of methanol and THF (1:1) (0.5 ml). N-NaOMe (0.1 ml) was added and stirred at room temperature for 2 hours. The mixture was concentrated. To the residue, there were added water (0.1 ml) and MeOH-THF (1:1) (0.5 ml) and stirred at room temperature for 2 hours. Amberlist A-15 (Tradename) was added to neutralize the mixture and then filtered. Amberlist A-15 was washed with methanol. The solution combined was concentrated in vacuo. The residue was washed with ether and dried to give the compound (22) (ganglioside GM 3 ) (1.7 mg).
• the compound (24) (216 mg, 0.195 mmol) was dissolved in 90% AcOH (3.0 ml). To this solution, there were added palladium chloride (41 mg) and sodium acetate (38 mg). The mixture was stirred for 5 hours in an ultrasonic stirrer, filtered and concentrated in vacuo. The residue was dissolved in ethyl acetate. The solution was washed with saturated NaHC0 3 solution and NaCl solution, dried on MgS0 4 and concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 10 g) and eluted with 4% MeOH-containing chloroform to give the compound (25) (117 mg, 56%).
• the compound (27) (27 mg, 0.015 mmol) was dissolved in methanol (0.5 ml). N-NaOMe (0.036 ml) was added to this solution and stirred at room temperature for 4 hours. THF (0.5 ml) was added to dissolve the crystals precipitated. The mixture was stirred for additional two hours and concentrated in vacuo. To the residue, there were added 80% MeOH solution (3.0 ml) in water and THF (3.0 ml). The mixture was stirred at room temperature for 5 hours and concentrated in vacuo. Water was added to the residue. Insolubles were collected to give the compound (28) (12.7 mg, 65.8%).
• Benzyl galactoside (the compound A) (8.1 g, 30 mmol) was suspended in acetone (150 ml). To this suspension, there were added paratoluenesulfonic acid (600 mg) and 2,2-dimethoxypropane (4.32 g). The mixture was stirred at room temperature for 2 hours. After triethylamine (2 ml) was added, the reaction mixture was concentrated in vacuo. The residue was subjected to silicagel column chromatography (Si02 C-300, 300 g) and eluted with toluene-ethyl acetate (1:2) to give the compound B (6.2 g, 66.7%).
• the compound G (5.11 g, 5.53 mmol) was dissolved in AcOH (50 ml), to which water (10 ml) was added. The mixture was stirred at 60°C for 2 hours and concentrated in vacuo. The resulting solid was suspended in and washed with hexane to obtain the compound H (4.82 g, 98.5%).
• the compound (31) (60 mg) was dissolved in MeOH (2.0 ml). N-NaOCH 3 (0.3 ml) was added. The mixture was stirred at room temperature for 24 hours, then neutralized by Amberlist A-15 (Tradename) and filtered. The filtrate was concentrated in vacuo to give the compound (34) as crystalline powder (33 mg, 67.8%).
• the compound (31) (200 mg) was dissolved in MeOH (12 ml), to which N-NaOMe (1.1 ml) was added. The mixture was stirred at room temperature for 24 hours, neutralized by Amberlite CG-50 (Tradename) and then filtered through Celite (Tradename). The residue was subjected to column chromatography (silanized silicagel, 10 g) and eluted with MeOH - H 2 0 (1:2) to give the compound (35) (109 mg, 63.6%).
• the compound (35) (108 mg) was dissolved in MeOH - H 2 0 (9:1) (5 ml). Catalytic reduction was carried out at room temperature for 24 hours and then at 60°C for 5 hours. The reaction mixture was concentrated in vacuo. The residue was subjected to Robar column (PR-8, size A) and eluted with MeOH - H 2 0 (80:1) to give the compound (36) (23.1 mg, 33.1%).
• the compound (32) (106 mg, 0.11 mmol) was dissolved in MeOH (3 ml), to which N-NaOMe (0.3 ml) was added. The mixture was stirred at room temperature for 24 hours, neutralized by Amberlist A-15 (tradename), and then concentrated in vacuo. The residue was subjected to Robar column (RP-18, size ⁇ ) and eluted with MeOH -H 2 0 (3:1) to give the compound (37) (60 mg, 70.5%).
• the compound (37) (63 mg) was dissolved in MeOH (2.0 ml), to which 10% Pd-C (63 mg) was added. Catalytic reduction was carried out for 24 hours. The reaction mixture was filtered to remove Pd-C and concentrated in vacuo to give almost quantitatively the compound (38).
• the compound (33) (260 mg, 0.28 mmol) was dissolved in MeOH (5 ml), to which N-NaOMe (1.12 ml) was added. The mixture was stirred at room temperature for 24 hours, neutralized by Amberlist A-15 (tradename), filtered and concentrated in vacuo. The residue was subjected to Robar column (RP-18, size ⁇ ) and eluted with MeOH - H 2 0 (3:1) to give the compound (39) (120 mg, 57.5%).
• the compound (39) (108 mg, 0.146 mmol) was dissolved in MeOH (5 ml), to which 10% Pd-C (200 mg) was added. Catalytic reduction was carried out for 24 hours. The mixture was filtered to remove Pd-C and concentrated in vacuo to give the compound (40) (68 mg, about 100%).
• the compound (32) (577 mg, 0.625 mmol) was dissolved in acetic anhydride (10 ml) and pyridine (10 ml). The mixture was stirred at room temperature for 24 hours. The reaction mixture was concentrated in vacuo. The residue was subjected to silicagel column chromatography (Si0 2 C-300, 80 g) and eluted with toluene-ethyl acetate (1:2) to give the compound (41) (477 mg, 79.1%).
• the compound (41) (379 mg, 0.39 mmol) was dissolved in MeOH (15 ml), to which 10% Pd-C (200 mg) was added. Catalytic reduction was carried out at room temperature for 24 hours. After filtration, the mixture was concentrated in vacuo. The residue was dissolved in a mixed solvent of acetic anhydride (5 ml) and pyridine (5 ml) and stirred at room temperature for 3 hours. The reaction mixture was concentrated in vacuo. The residue was dissolved in ethyl acetate, washed with water, dried on MgS0 4 and concentrated in vacuo. The residue was subjected to silicagel column chromatography (C-300, 20 g) and eluted with 15% MeOH - containing isopropylether to give the compound (42) (181 mg, 61.7%).
• the compound (33) (195 mg, 0.211 mmol) was dissolved in acetic anhydride (10 ml) and pyridine (10 ml). The mixture was stirred at room temperature for 24 hours and concentrated in vacuo. The residue was subjected to silicagel column and Robar column ( ⁇ -size) and eluted with toluene-ethylacetate (1:2) to give the compound (43) (148 mg, 72.6%).
• the compound (43) (99 mg, 0.10 mmol) was dissolved in MeOH (5 ml), to which 10% Pd-C (100 mg) was added. Catalytic reduction was carried out at room temperature for 24 hours. The reaction mixture was filtered to remove Pd-C and concentrated in vacuo. The residue was dissolved in acetic anhydride (1 ml) and pyridine (1 ml), stirred at room temperature for 24 hours and concentrated in vacuo. The residue was subjected to Robar column (size A) and eluted with ethyl acetate - toluene (2:1) to give quantitatively the compound (44) (77 mg).
• the compound (46) (136 mg, 0.1 mmol) was dissolved in MeOH (5 ml), to which N-NaOMe (0.8 ml) was added. The mixture was stirred at room temperature for 24 hours, neutralized by Amberlist A-15 (tradename), filtered to remove the resin and concentrated in vacuo. 0.1 N-NaOH solution (1.0 ml) was added to the residue. The mixture was stirred at room temperature for 24 hours, neutralized by Amberlist A-15 (tradename), filtered and concentrated in vacuo to give the compound (48) (96 mg, 80.6%).
• the compound (48) (84 mg, 0.071 mmol) was dissolved in MeOH (5 ml), to which 10% Pd-C (100 mg) was added. Catalytic reduction was carried out at room temperature for 2 days. The mixture was filtered to remove Pd-C and concentrated to give the compound (49) (42 mg, 93.7%).
• the compound (47) (136 mg, 0.1 mmol) was dissolved in MeOH (5 ml), to which N-NaOMe (0.6 ml) was added. The mixture was stirred at room temperature for 24 hours, neutralized by Amberlist A-15 (tradename), filtered and concentrated in vacuo. The residue was mixed with 0.1 N-NaOH solution (1.0 ml) and MeOH (2.0 ml) and stirred at room temperature for 7 hours. The reaction mixture was neutralized by Amberlist A-15 (tradename) and concentrated in vacuo to give the compound (50) (97 mg, 81.4%).
• the compound (47) (507 mg, 0.373 mmol) was dissolved in acetic anhydride (5.0 ml) and pyridine (5.0 ml) and stirred at room temperature for 24 hours.
• the reaction mixture was subjected to silicagel column chromatography (Wakogel C-300, 50 g) and eluted with 10% MeOH-containing toluene to give the compound (52) (484 mg, 93%).
• the compound (52) was dissolved in MeOH (15 ml), to which 10% Pd-C (200 mg) was added. Catalytic reduction was carried out. After the reduction was completed, the reaction mixture which contained the compound (53) (TLC, BuOH-EtOH-H 2 0 4:2:2, Rf 0.55) was filtered to remove Pd-C and concentrated in vacuo. The residue was dissolved in acetic anhydride (5 ml) and pyridine (5 ml) and stirred at room temperature for 2 hours. The mixture was concentrated in vacuo. The residue was subjected to silicagel column chromatography (C-300, 10 g) and eluted with ethyl acetate to give the compound (54) (235 mg, 69.3%).
• the compound (54) (190 mg, 0.171 mmol) was dissolved in DMF (1.0 ml) and heated to 50°C, to which hydrazinium acetate (19 mg) was added and stirred for 5 minutes. After cooled, there was added ethyl acetate (10 ml) to the reaction mixture and stirred for 30 minutes. The mixture was diluted with ethyl acetate and washed with saturated NaCl solution. The organic layer was dried on MgS0 4 and concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 10 g) and eluted with 1% MeOH-containing ethyl acetate to give the compound (19) (148 mg, 81.0%).
• the compound (19) (145 mg, 0.136 mmol) was dissolved in methylene chloride (1.0 ml). To this, there was added trichloroacetonitrile (54 ⁇ l) and NaH (60% in oil) (7.0 mg) under ice-cooled condition and stirred for 2 hours. The mixture was concentrated in vacuo. The residue was subjected to silicagel column chromatography (Wakogel C-300, 10 g) and eluted with ethyl acetate to give the compound (20) (110 mg, 66.8%).
• the compound (21) (25 mg, 0.014 mmol) was dissolved in methanol (0.5 ml). 1 N-NaOMe (56 ⁇ l) was added and stirred at room temperature for 24 hours. The mixture was concentrated. To the residue, there were added water (0.1 ml), THF (0.5 ml) and MeOH (0.5 ml) and stirred for 5 hours. Amberlist A-15 (Tradename) was added to neutralize the mixture and then filtered. The filtrate was concentrated in vacuo. The residue was recrystallized from MeOH to give the compound (22) (11 mg, 61.7%).

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• 1985
  • 1985-06-26 AU AU44212/85A patent/AU582758B2/en not_active Ceased
  • 1985-06-27 US US06/749,545 patent/US4968786A/en not_active Expired - Fee Related
  • 1985-06-27 ES ES544630A patent/ES8609354A1/es not_active Expired
  • 1985-06-27 HU HU852521A patent/HU196818B/hu not_active IP Right Cessation
  • 1985-06-27 EP EP85107982A patent/EP0166442A3/de not_active Ceased
  • 1985-06-28 CA CA000486085A patent/CA1262129A1/en active Granted
  • 1985-06-28 KR KR1019850004716A patent/KR920002826B1/ko not_active IP Right Cessation

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